Targeting glycolytic enzymes to combat cancer metastasis: Strategies toward clinical translation.

The Warburg effect, proposed a century ago, remains central to cancer metabolism research. It is characterized by the preference of cancer cells to convert glucose to lactate via glycolysis even under aerobic conditions. Metastasis is the leading cause of cancer-related mortality, and aerobic glycolysis promotes this process through multiple mechanisms, including enhancing anoikis resistance, inducing epithelial-mesenchymal transition (EMT), and promoting immunosuppression and angiogenesis. These pro-metastatic effects depend not only on the provision of energy and biosynthetic precursors but also on the non-metabolic moonlighting functions of glycolytic enzymes, making them promising therapeutic targets. This review systematically summarizes recent advances in the development and clinical translation of small-molecule inhibitors targeting glycolytic enzymes. In particular, we highlight the structural insights gained from co-crystal structures of glycolytic inhibitors bound to their enzymatic targets, which provide a molecular basis for rational drug design. It also outlines the principal strategies currently employed in their development and discusses key barriers to clinical application, including metabolic heterogeneity, compensatory mechanisms, and immunotoxicity. Importantly, mitochondrial signaling sustains tumor metabolic plasticity, enabling resistance to inhibition of a single metabolic target and highlighting mitochondria as promising targets for individualized cancer therapy. In the future, optimizing screening approaches, exploring combination therapies, and exploiting both the moonlighting functions of glycolytic enzymes and mitochondrial signaling may help overcome translational bottlenecks and accelerate the clinical application of anti-tumor metabolic therapies.